[Histonet] brain histology

[Cheryl Cross]

Hi Dr. Gibbons -

Great to see a fellow vet posting!! I do some neurotox stuff in mice.  
What makes for a really nice FFPE section is, if you can, perfuse  
fixing the rodents, then removing the calvarium overlying the brain,  
and letting the brain sit in the skull in your fixative of choice (I  
use 10% NBF). Letting the brain sit for 24 hours in the fix allows  
you to avoid those dreaded dark angular neurons. After sitting for 24  
hours I take out the brain and section routinely ... if I can't  
section immediately and immunohistochemistry will be performed i  
switch it to 70% EtOH ... but sitting in the alcohol makes the tissue  
very very firm. If immunohistochemistry isn't an issue I allow the  
brain or organs to remain in the 10% NBF.

Hope this helps, let me know if you need a protocol for perfusion, etc.

Cheryl

Cheryl Cross, DVM, Dipl. ACVP
Researcher
University Corporation for Atmospheric Research
College of Veterinary Medicine
University of Tennessee Department of Pathology
2407 River Drive, Room A201
Knoxville, TN 37996-4542
(423) 967-2724
fax: 865-974-5616
ccross <@t> ucar.edu




On May 9, 2007, at 4:50 PM, Rene J Buesa wrote:

> I think is too much time in alcohol(s). Storing in absolute EthOL  
> is not a good practice.
>   Try to reduce this aspect and check for results.
>   René J.
>
> I-sanna Gibbons <trinimaican2501 <@t> yahoo.com> wrote:
>   I am having a bit of trouble obtaining good quality slides of bat  
> brains. The brains are fixed in Carnoy's, stored in absolute  
> alcohol, processed routinely, infiltrated with paraffin, sectioned  
> at 8 microns and stained using cresyl fast violet. The aim is to  
> record the cytoarchitecture. The resulting slides show poor tissue  
> integrity with extensive cracking. Can anyone suggest how this can  
> be avoided? I was thinking maybe a double-embedding method for  
> whole brains (10-15 mm) and thicker sections (25-30 microns)?
>
> Thanking you in advance
>
> I-sanna Gibbons, DVM
> Veterinary Anatomy
> School of Veterinary Medicine
> The University of the West Indies
> Trinidad and Tobago, W.I.
>
>
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